[The regulation of retinoid X receptor-mediated oxidative stress pathway in rat pulmonary ischemia/reperfusion injury].

The aim of this study was to investigate the regulatory role of retinoid X receptor (RXR)-mediated oxidative stress pathway in rat pulmonary ischemia/reperfusion injury (PIRI) and the underlying mechanism. Seventy-seven male Sprague-Dawley (SD) rats were randomly divided into 7 groups (n = 11): control group, sham group, sham+9-cis-retinoid acid (9-cRA, RXR agonist) group, sham+HX531 (RXR inhibitor) group, ischemia/reperfusion (I/R) group, I/R+9-cRA group, and I/R+HX531 group. The unilateral lung I/R model was established by obstruction of left lung hilus for 30 min and reperfusion for 180 min in vivo. The rats in I/R+9-cRA and I/R+HX531 groups were given intraperitoneal injection of 9-cRA and HX531 before thoracotomy. After reperfusion, the left lung tissue was taken to evaluate the lung tissue injury, and the oxidative stress-related indexes of the lung tissue were detected by the corresponding kits. The lung tissue morphology and the ultrastructure of the alveolar epithelial cells were observed by HE staining and transmission electron microscope, respectively. The protein expression of RXR in lung tissue was observed by immunofluorescence labeling method, and the expression level of nuclear factor E2-related factor (Nrf2) protein was detected by Western blot. The results showed that, compared with the sham group, the I/R group exhibited obviously injured lung tissue, decreased SOD activity, increased MDA content and MPO activity, and down-regulated expression level of Nrf2 protein. Compared with the I/R group, the I/R+9-cRA group showed alleviated lung tissue injury, increased activity of SOD, decreased MDA content and MPO activity, and up-regulated expression levels of RXR and Nrf2 protein. The above-mentioned improvement effects of 9-cRA were reversed by HX531 treatment. These results suggest that RXR activation can effectively protect the lung tissue against I/R injury, and the mechanism may involve the activation of Nrf2 signaling pathway, the enhancement of antioxidant level and the reduction of oxidative stress response.

Isocitrate Dehydrogenase 2 Suppresses the Invasion of Hepatocellular Carcinoma Cells via Matrix Metalloproteinase 9.

BACKGROUND/AIMS: Isocitrate dehydrogenase 2 (IDH2) is a mitochondrial NADP-dependent isocitrate dehydrogenase, and has been found to be a tumor suppressor in several types of tumors. However, the roles of IDH2 in hepatocellular carcinoma (HCC) as well as underlying mechanisms remain unknown. METHODS: The IDH2 and matrix metalloproteinase 9 (MMP9) levels in the specimens from 24 HCC patients were investigated by Western blot and ELISA, respectively. Their relationship was examined by correlation analyses. Patient survival with high IDH2 levels and low IDH2 levels was compared. IDH2 levels and MMP9 levels were modified in a human HCC cell line. The effects of IDH2 or MMP9 modulation on the expression of the other were analyzed. The effects of IDH2 on cell invasion were analyzed in a transwell cell invasion assay. The dependence of nuclear factor x03BA;B (NF-x03BA;B) signaling was examined using a specific inhibitor. RESULTS: The IDH2 levels significantly decreased in HCC, and were lower in HCC with metastases, compared to those without metastases. IDH2 levels inversely correlated with MMP9 levels in HCC. HCC patients with Low IDH2 had lower 5-year survival. MMP9 levels did not regulate IDH2 levels, while IDH2 inhibited MMP9 levels in HCC cells, in a NF-x03BA;B signaling dependent manner, possibly through ix03BA;B, to suppress HCC cell invasion. CONCLUSIONS: Down regulation of IDH2 may promote HCC cell invasion via NF-x03BA;B-dependent increases in MMP9 activity. IDH2 may be a potential therapeutic target for HCC.

Non-alcoholic fatty liver disease (NAFLD) fibrosis score predicts 6.6-year overall mortality of Chinese patients with NAFLD.

The non-alcoholic fatty liver disease (NAFLD) fibrosis score (NFS) has emerged as a useful predictor of long-term outcome in NAFLD patients. We evaluated the predictive performance of the NFS for overall mortality in a Chinese population with NAFLD. All NAFLD patients diagnosed ultrasonographically at Xixi Hospital of Hangzhou between 1996 and 2011 were retrospectively recruited to the study. Outcome was determined by interview and causes of death were confirmed by medical records. The area under the receiver operating characteristic curve (AUCROC ) was used to determine the predictive accuracy of the NFS, BARD (body mass index, aspartate aminotransferase (AST)/alanine aminotransferase (ALT) ratio, diabetes) score, FIB-4 index and the AST/platelet ratio index (APRI) for mortality. Data from a total of 180 eligible patients (median age 39 years; 96 men) were analysed, with 12 deaths over a median follow-up period of 6.6 years (range 0.5-14.8 years). Using Cox model analysis, the NFS as a continuous variable was identified as the only predictor for all-cause mortality (hazard ratio 2.743, 95% confidence interval (CI) 1.670-4.504). The NFS yielded the highest AUCROC of 0.828 (95% CI 0.728-0.928, P < 0.05), followed by the FIB-4 index, APRI and BARD score (AUCROC 0.806 (P < 0.05), 0.732 (P < 0.05) and 0.632, respectively). The data indicated that the NFS is a useful predictor of 6.6-year all-cause mortality for Chinese patients with NAFLD.

Toll-like receptor-4 signalling in the progression of non-alcoholic fatty liver disease induced by high-fat and high-fructose diet in mice.

The aim of the present study was to investigate Toll-like receptor-4 (TLR4) signalling at different stages of non-alcoholic fatty liver disease (NAFLD) induced by a high-fat, high-fructose (HFHFr) diet in mice. Both TLR4 wild-type (WT) and mutant (TLR4(mut) ) mice were fed either standard chow (SC) or the HFHFr diet for different periods of time from 4 to 16 weeks. Pathological characteristics and function of the liver were assessed. Simple steatosis, steatohepatitis and hepatic fibrosis occurred sequentially in Week 4, 8 and 16 in WT mice fed with the HFHFr. Expression of TLR4, myeloid differentiation factor 88 (MyD88), interferon regulatory factor (IRF) 3 and IRF7 started to increase at Week 4, peaked at Week 8 and then declined to basal levels at Week 16. This pattern was consistent with changes in inflammation in the liver revealed by haematoxylin and eosin staining. However, lipid accumulation, inflammation and fibrosis in livers of TLR4(mut) mice fed the HFHFr diet were significantly alleviated. In addition, the expression of activin A in WT mice fed the HFHFr diet increased at Week 16. The data suggest that TLR4 signalling mediates non-alcoholic steatohepatitis before fibrosis and that activin A is subsequently involved in NAFLD.

Detection of hepatitis C virus by an improved loop-mediated isothermal amplification assay.

An improved, sensitive, specific, and rapid one-step reverse transcription loop-mediated isothermal amplification (LAMP) assay targeting the 5' untranslated region (UTR) was developed to detect hepatitis C virus (HCV) infection. Based on an accelerating primer (AP), the present assay, named AP-LAMP, has the advantages of rapidity and sensitivity over the routine LAMP method. The possible AP-based amplification pathway during the reaction was revealed by restriction enzyme digestion and eletrophoresis. The detection limit of the AP-LAMP assay was approximately 84 IU/ml, and no cross-detection was observed. The assay was evaluated further with 126 clinical specimens, and the results indicated the suitability and simplicity of the test as a rapid diagnostic tool for detection of HCV RNA.

Rapid identification of YVDD mutants of hepatitis B virus using smart amplification process with competition probe.

Accurate and timely detection of drug-associated viral mutants is important during antiviral therapy. Combining Smart Amplification Process (SMAP) with competition probe, an assay specifically designed to detect point mutation at codon 204 of the hepatitis B virus (HBV) polymerase gene was developed. This assay was sensitive to detect 20 copies of mutant/reaction and recognize as little as 1% of minor mutants in the viral population. The comparison of direct sequencing and SMAP method on 35 clinical specimens showed the concordance in 88% of the cases. This method provides an efficient alternative for rapid identification of HBV mutation associated with lamivudine resistance.

[Efficacy and safety of lamivudine plus adefovir combination therapy and entecavir monotherapy for chronic hepatitis B patients].

To compare the efficacy and safety of Lamivudine (LAM) plus Adefovir dipivoxil (ADV) combination therapy and Entecavir (ETV) monotherapy for chronic hepatitis B patients. 120 patients with chronic hepatitis B managed in a single-centre clinical practice (median 96 weeks) were split into 2 cohorts, one was treated with de-novo combination Lamivudine (100 mg/day) plus Adefovir (10 mg/day) (LAM+ADV), the other with Entecavir (0.5 mg/day) monotherapy. Serum levels of ALT, creatinine, HBsAg, HBeAg and HBV viral load, together with genotypic resistence were analyzed at 0, 12, 24, 48, 96 weeks, respectively. HBV DNA was determined by real-time PCR. HBsAg and HBeAg were assessed by chemiluminescence. Serum levels of ALT and creatinine were detected by automatic biochemical analyzer. HBV genotypic resistence was tested by direct sequencing. (1) At the time point of 96 weeks, a total of 99 patients (51 cases in combination therapy cohort and 48 case in monotherapy cohort) were compared. The baseline characteristics as for HBV viral load, median age, serum levels of ALT and creatinine were compatible between combination therapy cohort and monotherapy cohort. (2) The rates of HBV DNA values is less than 300 copies/ml and HBV DNA values is less than 1000 copies/ml had no significant difference between LAM + ADV and ETV cohorts by the 12 and 24 weeks (P more than 0.05). (3) At the time point of 48 weeks, the rates of HBV DNA is less than 1000 copies/ml, HBeAg seroconversion, and ALT normalization were similar in both cohorts, though the rate of HBV DNA values is less than 300 copies/ml was obviously higher in combination therapy cohort than that of monotherapy cohort (90.7% vs 76%, P values is less than 0.05). (4) At the time point of 96 weeks, the rates of HBV DNA values is less than 300 copies/ml (96.1% vs 79.2%), HBV DNA values is less than 1000 copies/ml (98% vs 87.5%) and the HBeAg seroconversion (41.7% vs 16.7%) were markedly higher in combination therapy cohort than those of monotherapy cohort statistically (P values is less than 0.05 for all). The mean values of decreases for HBV viral loads and HBsAg levels were smilar in both cohorts at 48 and 96 weeks. (5) Elevated serum creatinine not be found in both cohorts at the end of treatment. (6) No virological breakthrough occurred in combination therapy cohort at the end of treatment. Four patients in monotherapy cohort were found with virological breakthrough at 96 weeks and three cases among were confirmed to be of variants associated with ETV resistance (rtL180M + T184L + M204V). Present study suggests that Lamivudine plus Adefovir dipivoxil de-novo combination therapy was more efficacious than Entecavir monotherapy for CHB patients and the tolerance is compatible.

[Determination of ascitic bacterial 16S rRNA by quantitative PCR-microarray in the diagnosis of spontaneous bacterial peritonitis].

OBJECTIVE: To evaluate the significance of determining ascitic bacterial 16S rRNA by quantitative PCR combined with microarray (PCR-microarray) in the diagnosis of spontaneous bacterial peritonitis (SBP). METHODS: Ascitic bacterial 16SrRNA was determined by real time fluorescent quantitative PCR-microarray in 76 cases of suspected SBP and 6 cases of non-infectious ascites with chronic liver diseases. The results were compared with ascitic bacterial culture simultaneously. RESULTS: Of 76 ascitic samples, 17 were detected bacteria positive by PCR-microarray, including 8 Grams positive(G+) and 9 Grams negative(G-), which was higher than that by bacterial culture which had only 6 ascitic samples detected positive (all G-); the positive rates were 22.4% vs 7.9%, respectively (P < 0.01). The bacterial strains detected by both methods in 6 cases had a consistency with each other. No bacteria were detected in another 6 cases of non-infectious ascites with chronic liver diseases. CONCLUSIONS: Determination of ascitic bacteria 16S rRNA by PCR-microarray has a higher specificity and sensitivity in the diagnosis of SBP as compared with the bacteria culture. Application of this novel method can not only accelerate SBP diagnosis but also stratify the different pathogens.

[The regulatory role of autophagy in rats lung ischemia/reperfusion injury].

Objective: To investigate the role of cell autophagy in lung ischemia/reperfusion injury in rats. Methods: Forty SD rats were randomly divided into 5 groups (n=8): ①Sham operated group (sham group):just open rat chest for 3.5 h; ②Ischemia/reperfusion group (I/R group):after open chest, clamp pulmonary hilus for 0.5h then reperfusion for 3 h; ③Solvent group (DMSO group): intraperitoneal injection of DMSO solution for 1h before operation; ④Autophagic inhibitor group (3-MA group); ⑤Autophagic agonist group (Rap group): intraperitoneal injection of autophagic agonist rapamycin before operation; the rest operations of DMSO, 3-MA and Rap groups are the same as that of I/R group. At the end of the experiment, the rats were killed by euthanasia-killing. The lung tissues were collected and the wet/dry weight ratio (W/D) and total lung water content (TLW) of the lung tissues were detected. The lung tissue structure and cell ultramicro morphology were observed by light microscopy and electron microscopy and the injuried alveolar rate(IAR) was calculated. The autophagy-related protein expressions were detected by Western blot. Results: Compared with sham group, the levels of W/D, TLW and IAR were increased, the expressions of autophagy related protein and p-AMPK, Beclin 1, LC3 II were also increased in other four groups, while the protein expressions of p-mTOR and p62 were decreased significantly (P＜ 0.05 or P＜0.01). Under the light microscope, the other groups of lung tissue had edema and exudation in varying degrees, the structure of alveoli was disordered, the ultrastructural damage of cells was aggravated under the electron microscope, and autophagosome could be observed. Compared with DMSO group, the expressions of autophagy related protein, the levels of W/D, TLW and IAR in 3-MA group were decreased (P＜0.05 or P＜0.01), the edema of lung interstitial was lighter, and less cells were found in alveolar cavity. Ultrastructural damage was also lighter and with less autophagosome. Besides, there was no significant difference among I/R, DMSO and Rap groups (P＞0.05). Conclusion: Autophagy can be activated during ischemia/reperfusion in rats to induce lung injury.

[Determination of ascitic bacterial 16S rRNA gene in the rapid diagnosis of spontaneous bacterial peritonitis].

OBJECTIVE: To evaluate the value of ascitic bacterial 16S rRNA gene determination in the rapid diagnosis of spontaneous bacterial peritonitis (SBP). METHODS: 16S rRNA gene from bacterial DNA in ascites was determined by quantitative fluorescent polymerase chain reaction (PCR) in 76 patients with suspected SBP and 6 patients with non-infectious ascites. The results were compared with those obtained from bacterial culture. RESULTS: The positive rate of SBP was 22.4% among patients detected with ascitic bacterial 16S rRNA gene determination-based quantitative fluorescent PCR, which was significantly higher than that (7.9%) in patients only received bacterial culture (P<0.05). In addition,in 6 patients with non-infectious ascites,both the 16S rRNA gene determination-based quantitative fluorescent PCR and bacterial culture showed negative results. CONCLUSIONS: 16S rRNA gene determination-based quantitative fluorescent PCR can be an effective tool for the rapid diagnosis of SBP. It is more sensitive than the bacterial culture.

Anti-hepatitis B virus treatment guided by long-term virus kinetics stepwise tracing.

BACKGROUND AND AIMS: Biphasic modeling of viral kinetics provided valuable information to rapidly assess potential antiviral regimens during the beginning of therapy, but has not been performed over the long term. METHODS: In 11 chronic hepatitis B patients with lamivudine treated per day, the model with delay time was applied to examine phase transition and analyze viral kinetics parameters. RESULTS: Viral decay conformed to a biphasic mode during the first 4 weeks, with the complex profile appeared in the later period. Lamivudine treatment resulted in a mean log hepatitis B virus (HBV)-DNA decline of 1.77 +/- 0.55 after 4 weeks and 3.79 +/- 1.70 log after 24 weeks. The median effectiveness of blocking viral replication was 96% (range, 89-99%). The median rate of free virus clearance and infected cell loss was 1.1/day and 0.03/day, respectively. Through phase transition determination and stepwise modeling process, viral kinetics were evaluated for complex decay profile during long-term therapy. Moreover, with the abnormal kinetics tracked, an occasion of add-on combined therapy was developed to treat patients with emerging virus mutants. CONCLUSION: The present study using mathematical modeling of viral decay may be a useful approach to evaluate optimal individualized therapy for HBV infection in a continuous long-term manner.

[Clinical and histological features of non-alcoholic fatty liver disease].

OBJECTIVE: To investigate the clinical and histological features in Chinese patients with non-alcoholic fatty liver disease (NAFLD). METHODS: 108 patients with biopsy-proven NAFLD were enrolled in this study. Clinical, demographic, and biochemical data were compared between NAFLD patients with abnormal ALT and those with normal ALT. RESULTS: Simple fatty liver, nonalcoholic steatohepatitis(NASH) and cirrhosis were diagnosed in 49 (45.4%), 57(52.7%) and 2 (1.9%) patients, respectively. ALT and AST levels of NASH group were higher than those of simple fatty liver group (t = 2.55, 3.13; P = 0.01, 0.00). Fifty of the 77 patients (64.9%) with abnormal ALT levels were diagnosed as non-alcoholic steatohepatitis (NASH), and twenty-six were diagnosed as simple fatty liver, according to liver histology. Among the 31 patients with normal ALT levels, nine (29%) had NASH and twenty-two had simple fatty liver (P = 0.00). The patients with normal ALT had lower necroinflammatory grade than patients with abnormal ALT (x2 = 10.30, P = 0.01), but they had similar degree of steatosis and fibrosis (x2 = 5.52, 6.12; P = 0.12, 0.01). AST, g-glutamyltransferase, total cholesterol, apolipoprotein A1, apolipoprotein B and systolic blood pressure of patients with normal ALT were all lower than those of patients with abnormal ALT (t = 5.91, 2.00, 2.30, 2.10, 3.14, 2.43; P = 0.00, 0.05, 0.02, 0.04, 0.00, 0.02), while spleen thickness and AST/ALT ratio in patients with normal ALT were higher than those with abnormal ALT significantly (t = 3.70, 2.95; P = 0.00, 0.01). Multivariate analysis revealed that ALT (OR = 2.78, 95% CI 1.06-7.3, P = 0.04) was the only independent predictor of NASH, and ALT had low accuracy in predicting NASH, the area under the receiver operating characteristics curves of ALT to predict NASH was 0.69 (95% CI 0.59-0.8, P = 0.00). CONCLUSION: NAFLD patients have higher ALT level, and elevated serum level of ALT is independent predictor of the degree of inflammation, but not of steatosis and fibrosis.

[The roles of saliva testing for preventing hepatitis B virus spreading].

OBJECTIVE: To discuss the significance of testing hepatitis B virus (HBV) from saliva in HBV patients. METHODS: HBV DNA content in serum and saliva of 200 HBV patients and 20 healthy subjects were detected by fluorescence quantitative polymerase chain reaction. According to the serum level of HBV content, four groups were divided: control group A, group B negative, low virus C (1 x 10(3) - 1 x 10(5) copies/ml) and high-group D ( > 1 x 10(5) copies/ml). The relationship of serum and virus content in saliva was analysed. RESULTS: Of 200 HBV cases, 180 were found HBV DNA in serum with positive rate of 90.0%; while 145 were found HBV DNA in saliva with positive rate of 72.5%, and there was no significant difference (chi2 = 1.35, P > 0.05). The significant difference was observed in testing serum and saliva in Group C (100.0% vs. 38.5%; Z = 14.11, P < 0.01). In group D, there was no significant difference found either (100.0% vs. 83.8%; chi2 = 1.05, P > 0.05). Group D virus serum had a high average level of (6.63 +/- 1.55) log copies/ml virus and in the saliva had an average level of (5.21 +/- 1.85) log copies/ml; saliva had serum viral load lower than an order of magnitude average. No HBV DNA was found in serum or saliva from 20 health subjects. CONCLUSION: When the serum contains a high content of HBV DNA virus, the content of saliva HBV DNA virus should be likely high, which might pose a threat of source of infection. A precise quantitative detection of HBV DNA in saliva might be used as evaluation of the level of virus in the body copy for judgment of infection.

[Clinical significance of intrahepatic hepatitis B core antigen (+) in patients with chronic hepatitis B].

OBJECTIVE: This study aimed to assess the clinical significance of intrahepatic hepatitis B core antigen (HBcAg) (+) in patients with chronic hepatitis B (CHB). METHODS: 200 CHB patients were prospectively studied using fluorescence quantitative PCR (FQ-PCR), combined PCR with fluorescence probe hybridization technique, to determine serum HBV DNA. Serum HBeAg was measured quantitatively. Liver biopsies were performed and immunohistochemistry stained liver slides were examined in all the cases. Correlation analyses were performed. RESULTS: Based on the HBV DNA levels, the patients were divided into 5 groups: group A (<3 log10 copies/ml) n=20, group B (>or=3 log10 copies/ml-<5 log10 copies/ml) n=13, group C (>or=5 log10 copies/ml-<6 log10 copies/ml) n=24, group D (>or=6 log10 copies/ml-<8 log10 copies/ml) n=116, and group E (>or=8 log10 copies/ml) n=27, and 87.5% of the CHB patients were intrahepatic HBcAg (+). The rate of HBcAg (+) was 55.0% (11/20) in group A, 53.8% (7/13) in group B, 75.0% (19/24) in group C, 96.6% (112/116) in group D, and 100% (27/27) in group E. A strong correlation was found between the rate of HBcAg (+) and the level of serum HBV DNA (r=0.80). This type of association also appeared between serum HBV DNA levels and HBeAg (+) (r=0.47). Of 20 CHB patients who were serum HBV DNA negative, 25% (5) were HBeAg (+), and 55% (11) were HBcAg (+), whereas 15 patients were both HBV DNA (-) and HBeAg (-), and 46.7% (7) were HBcAg (+). CONCLUSIONS: Intrahepatic HBcAg (+) in CHB patients might be more reliable in reflecting HBV replication. Determination of HBcAg (+) may have clinical significance for evaluating the efficacy of antiviral therapy and for predicting the therapeutic responses to different antiviral agents.

Baseline HBsAg predicts response to pegylated interferon-α2b in HBeAg-positive chronic hepatitis B patients.

AIM: To evaluate the predictive effect of baseline hepatitis B surface antigen (HBsAg) on response to pegylated interferon (PEG-IFN)-α2b in hepatitis B e antigen (HBeAg)-positive chronic hepatitis B (CHB) patients. METHODS: This retrospective analysis compared the treatment efficacy of PEG-IFN-α2b alone in 55 HBeAg-positive CHB patients with different baseline HBsAg levels. Serum HBV DNA load was measured at baseline, and at 12, 24 and 48 wk of therapy. Virological response was defined as HBV DNA < 1000 IU/mL. Serum HBsAg titers were quantitatively assayed at baseline, and at 12 and 24 wk. RESULTS: Eighteen patients had baseline HBsAg > 20 000 IU/mL, 26 patients had 1500-20000 IU/mL, and 11 patients had < 1500 IU/mL. Three (16.7%), 11 (42.3%) and seven (63.6%) patients in each group achieved a virological response at week 48, with a significant difference between groups with baseline HBsAg levels > 20000 or < 20000 IU/mL (P = 0.02). Thirteen patients had an HBsAg decline > 0.5 log10 and 30 patients < 0.5 log10 at week 12; and 6 (46.2%) and 10 (33.3%) in each group achieved virological response at week 48, with no significant difference between the two groups (P = 0.502). Eighteen patients had an HBsAg decline > 1.0 log10 and 30 patients < 1.0 log10 at week 24, and 8 (44.4%) and 11 (36.7%) achieved a virological response at week 48, with no significant difference between the two groups (P = 0.762). None of the 16 patients with HBsAg > 20000 IU/mL at week 24 achieved a virological response at week 48. CONCLUSION: Baseline HBsAg level in combination with HBV DNA may become an effective predictor for guiding optimal therapy with PEG-IFN-α2b against HBeAg-positive CHB.

[The relationship between serum HBsAg levels and liver inflammation and fibrosis in patients with chronic hepatitis B].

OBJECTIVE: To investigate the relation of hepatitis B surface antigen (HBsAg) level with chronic hepatitis B (CHB) and liver inflammation and fibrosis. METHODS: A total of 301 patients who diagnosed CHB and underwent liver biopsy were enrolled into the study. Meantimes, the biochemical markers, ferritin (FERR), serum HBsAg and HBV DNA quantitation were detected. The relation between HBsAg level and liver pathology were determined by spearman rank correlation analysis. The receiver operating characteristic curve was used to evaluate the accuracy of HBsAg level for liver inflammation and fibrosis. RESULTS: The body mass index (BMI), age, gender, genotype and family history had no effective on liver inflammation and fibrosis (P < 0.05). With the progressing of inflammation and fibrosis, the serum AST and ALT raise obviously (chi2 = 71.193, 96.344, 47.847, 63.981; P = 0.000, 0.000, 0.000, 0.000). When fibrosis reached to S4, the level of HBV DNA decreased obviously (chi2 = 33. 322; P = 0.000). With the aggravation of inflammation and fibrosis, the serum HBsAg gradually descended (chi2 = 68.173,15.719; P = 0.000, 0.000). The areas under operating characteristics curves of HBsAg predicted < or = G3 and < or = S3 were 0.732 and 0.793, and the specificity were 0.778, 0.891, and sensitivity were 0.685, and 0.633, respectively. CONCLUSION: The level of HBsAg of Chinese CHB patients descended gradually with the aggravation of liver inflammation and fibrosis. The serum HBsAg had a higher specificity to predict < or = G3 and < or = S3 of CHB patients. But there had superiority of predicting fibrosis than inflammation.

[Genotypes and evolution characteristics of three patients with poor response to initial treatment of Lamivudine and Adefovir dipivoxil for hepatitis].

OBJECTIVE: To understand the genotype characteristics and its evolution of patients with poor response to initial combined treatment of Lamivudine and Adefovir dipivoxil for chronic hepatitis B. METHODS: We detected the HBV genotypes of three patients-S1, S2, S3, who with poor response to initial treatment of Lamivudine and Adefovir dipivoxil for chronic hepatitis B over 12 months by the application of cloning and sequencing method at the time point of baseline,4 weeks after treatment, 12 weeks, 24 weeks, 48 weeks, 60 weeks. 25 clones were randomly selected to identify and sequence at each time point. RESULTS: The total number of clones from 3 patients with poor response to initial combined treatment of Lamivudine and Adefovir dipivoxil for chronic hepatitis B at each time point was 398. About patient S1 at baseline, genotype C accounting for 8.3%, genotype B, for 91.7%, so genotype B was in dominant (22/24). But genotype C has gradually developed to 100% after treatment for 60 weeks. About patient S2 and S3, genotype B was the only type at baseline. However type B has gradually "drift" to type C during treatment. When treatment for 60 weeks, type C has taken the absolute advantage 75% for S2, and 100% for S3. CONCLUSIONS: The cloning and sequencing can represent the overall genotype level better. HBV genotype has performed the evolution trend that genotype has drifted from B to C during long-term drug pressure, which is the main reason for poor response to initial combined treatment of Lamivudine and Adefovir dipivoxil for chronic hepatitis B.

[Three-year efficacy and side effect of adefovir dipivoxil for the treatment of the old patients with chronic hepatitis B virus infection].

OBJECTIVE: To investigate 3-year antiviral efficacy and side effect of adefovir dipivoxil (ADV) on the old patients with hepatitis B chronic infection. METHODS: 31 HBeAg-negative chronic hepatitis B virus infected old patients (include 8 patients with chronic hepatitis B and 23 patients with liver cirrhosis) with serum HBV DNA levels > 1000 copies/ml, and ALT > 2 times the upper limit of normal, without company with other liver diseases, cancer, renal dysfunction, and autoimmune disease. All the patients were treated with ADV orally (10 mg once daily) for 36 months. HBV DNA and biochemical and blood routine indexes were checked after treated. RESULT: Serum total bilirubin, direct bilirubin, alamine aminotransferase, aspartate aminotransferase and load of HBV DNA decrease significantly after therapy (P < 0. 001). Other biochemical indexs and blood routine are no significant changes (P > 0.05). CONCLUSION: The way to treat with ADV is safe and effective for old patients with chronic hepatitis B virus infection.